Presenter/Author(s) Information

Chandler CookFollow
Rudy CastilloFollow
Lisa HodgeFollow

Departmental Affiliation and City, State, Zip for All Authors

Department of Anatomy and Physiology University of North Texas Health and Science 3500 Camp Bowie Fort Worth, Tx 76107

Scientific Abstract

Lipopolysaccharide (LPS) is an endotoxin found in gram-negative bacteria such as Escherichia coli (E. coli). LPS protects the bacterial cell membrane; however, if the bacteria dies, LPS is released into the tissue where it activates the innate immune system and triggers an inflammatory responses. Macrophages are large phagocytes can that engulf pathogens, activate leukocytes and release inflammatory mediators such as nitrite (NO2-) and cytokines. The purpose of this study was to measure the macrophage inflammatory response against LPS in vitro. We hypothesized that culturing macrophages with LPS would increase the production of nitrite (NO2) and the pro-inflammatory cytokine, tumor necrosis factor alpha (TNF-). In these experiments, murine RAW 264.7 cells were grown in Dulbecco’s modified eagle high glucose medium with 10% fetal bovine serum at 37oC under 5% CO2. Cells were cultured for one hour to allow cells to acclimate. After one hour, LPS was added (500ng/well) and the cells were cultured for 24 hours. Cell-free supernatants were assayed for nitrite (NO2-) using the Griess reagent. The concentration of TNF-was measures using an ELISA. LPS increased the release of inflammatory mediators from RAW 264.7 macrophages. Specifically, LPS increased the production of NO2- approximately 7-fold and TNF- approximately 17-fold. In future studies, in vitro experiments will be used to test the inhibitory effect of anti-inflammatory therapeutics on macrophage activity.

Share

COinS
 

MACROPHAGES PRODUCE AN INFLAMMATORY RESPONSE TO LIPOPOLYSACCHARIDE (LPS)

Lipopolysaccharide (LPS) is an endotoxin found in gram-negative bacteria such as Escherichia coli (E. coli). LPS protects the bacterial cell membrane; however, if the bacteria dies, LPS is released into the tissue where it activates the innate immune system and triggers an inflammatory responses. Macrophages are large phagocytes can that engulf pathogens, activate leukocytes and release inflammatory mediators such as nitrite (NO2-) and cytokines. The purpose of this study was to measure the macrophage inflammatory response against LPS in vitro. We hypothesized that culturing macrophages with LPS would increase the production of nitrite (NO2) and the pro-inflammatory cytokine, tumor necrosis factor alpha (TNF-). In these experiments, murine RAW 264.7 cells were grown in Dulbecco’s modified eagle high glucose medium with 10% fetal bovine serum at 37oC under 5% CO2. Cells were cultured for one hour to allow cells to acclimate. After one hour, LPS was added (500ng/well) and the cells were cultured for 24 hours. Cell-free supernatants were assayed for nitrite (NO2-) using the Griess reagent. The concentration of TNF-was measures using an ELISA. LPS increased the release of inflammatory mediators from RAW 264.7 macrophages. Specifically, LPS increased the production of NO2- approximately 7-fold and TNF- approximately 17-fold. In future studies, in vitro experiments will be used to test the inhibitory effect of anti-inflammatory therapeutics on macrophage activity.

Manuscript Number

1030